Stabilization of the Reductase Domain in the Catalytically Self-Sufficient Cytochrome P450_BM3 by Consensus-Guided Mutagenesis.

ABSTRACT: The multidomain, catalytically self-sufficient cytochrome P450 BM-3 from Bacillus megaterium (P450_BM3 ) constitutes a versatile enzyme for the oxyfunctionalization of organic molecules and natural products. However, the limited stability of the diflavin reductase domain limits the utility of this enzyme for synthetic applications. In this work, a consensus-guided mutagenesis approach was applied to enhance the thermal stability of the reductase domain of P450_BM3 . Upon phylogenetic analysis of a set of distantly related P450s (>38 % identity), a total of 14 amino acid substitutions were identified and evaluated in terms of their stabilizing effects relative to the wild-type reductase domain. Recombination of the six most stabilizing mutations generated two thermostabl

SUBMITTER: Saab-Rincon G 

PROVIDER: S-EPMC5941085 | biostudies-literature | 2018 Mar

REPOSITORIES: biostudies-literature